The Platform Protein Is Essential for Type IV Pilus Biogenesis

• DOI: 10.1074/jbc.m113.453506
• Publication Date: 2013-02-15T01:15:40Z
• Authors: Herlinder K. Takhar; Kevin Kemp; Melissa Kim; P.Lynne Howell; Lori L. Burrows
• Keywords: Adenosine Triphosphatases; Models, Molecular; 0301 basic medicine; Cytoplasm; Polymers; Protein Conformation; Surface Properties; Molecular Motor Proteins; Cell Membrane; Membrane Proteins; Proteins; Bacterial Adhesion; Protein Structure, Tertiary; 03 medical and health sciences; Bacterial Proteins; Fimbriae, Bacterial; Pseudomonas aeruginosa; Mutagenesis, Site-Directed; Fimbriae Proteins; Plasmids; Protein Binding

A systematic genetic analysis was performed to identify the inner membrane proteins essential for type IV pilus (T4P) expression in Pseudomonas aeruginosa. By inactivating retraction aspect of function, genes T4P assembly were discriminated. In contrast previous studies system Neisseria spp., we found that components subcomplex consisting PilMNOP not surface expression, whereas highly conserved protein PilC essential. Here, present data may coordinate activity cytoplasmic polymerization (PilB) and depolymerization (PilT) ATPases via their interactions with its two domains. Using vitro co-affinity purification, show PilB interacts N-terminal domain PilC. We hypothesized PilT similarly C-terminal domain. Overexpression wild-type reduced twitching motility by ∼50% compared vector control. Site-directed mutagenesis T4P-specific residues yielded mutant supported but had a capacity support motility, suggesting impairment putative PilC-PilT interactions. Taken together, our results is an component system, controlling both disassembly.